MORPHOLOGICAL MECHANISMS OF ORGANELLE FUNCTION AND TRANSFORMATION IN CULTURE CELL

培养细胞中细胞器功能和转化的形态机制

• 批准号: 3939274
• 负责人: M C WILLINGHAM
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3939274
• 关键词: Golgi apparatus; antitumor antibody; cellular oncology; daunorubicin; drug resistance; electron microscopy; exocytosis; gel electrophoresis; genetic translation; histochemistry /cytochemistry; human tissue; hybridomas; immunochemistry; immunoconjugates; immunofluorescence technique; immunoperoxidase; lysosomes; microscopy; molecular biology; monoclonal antibody; neoplasm /cancer immunotherapy; neoplastic transformation; oncoproteins; organelles; ovary neoplasms; tissue /cell culture; tissue /cell preparation; tumor antigens

Neoplastic transformation produces many changes in cell physiology, some of which can be studied using morphologic techniques. We have used morphologic methods to study three general areas: 1) We have employed light microscopic immunocytochemistry to evaluate and select monoclonal antibodies for use as immunotoxin reagents for cancer therapy. A large scale screening chamber has been devised for use with immunofluorescence as a primary selection for surface-reactive monoclonal antibodies to human ovarian cancer cells. Hybridoma clones selected in this way were further evaluated by screening against normal human tissues and ovarian tumors using peroxidase immunocytochemistry in cryostat sections. Using these methods we have found two hybridomas highly reactive with human ovarian tumors: OVB1 and OVB3. We have characterized the tissue distribution and biochemical properties of these antigens in cultured cells, in tumors and in normal tissues. 2) We have studied the phenomenon of multidrug-resistance (MDR) in cultured cells, and have used a monoclonal antibody (MRK-16) to the human mdrl gene product (P170) as a probe to demonstrate P170 on the surface and in the Golgi stacks of drug-resistant KB cells in culture. We also showed the absence of P170 from coated pits, suggesting that this protein is immobilized in the plasma membrane. We have also used this antibody to examine the distribution of P170 in normal human tissues, and we have evaluated the amounts of P170 in progressively resistant derivatives of KB cells using immunofluorescence. 3) In other studies, we have localized p55, a major thyroid hormone binding protein found in the endoplasmic reticulum and nuclear envelope of cultured cells in normal tissues using immunoperoxidase histochemistry. This protein was found in highest amounts in cells that have abundant endoplasmic reticulum. Other cytochemical studies demonstrated a role for clathrin-coated pits in the loss of ADH-responsive water transport in isolated perfused rabbit cortical collecting ducts. We also demonstrated the localization of carbohydrate sites reactive with wheat germ lectin on nuclear pores in isolated nuclei.

肿瘤转化使细胞发生许多变化